Background: Esophageal cancer is one of the most common cancer worldwide with poor prognosis and high mortality. SNAI1 transcription factors, coding Snail1, it is important for the development of esophageal cancer metastasis while microRNA (miRNA) -203 has been shown to function as inhibitors of metastasis in the EC. The EC Snail1 protein is stabilized in part by deubiquitinating enzymes USP26; However, how USP26 set is not fully known.
Methods: Expression SNAI1 and USP26 messenger RNA (mRNA) and miR-203 performed on datasets in The Cancer Genome Atlas and Gene Expression Omnibus, respectively. Snail1 and USP26 protein expression and miR-203 are determined in normal esophageal cell lines HET-1A and Kyse150 EC cell lines and TE-1 using western blot and quantitative polymerase chain reaction, respectively. TargetScan used for in situ predicted miR-203 targets and in vitro assays using a heterologous reporter wild-type and miR-203 mutant seeds from the 3 ‘translated region (UTR) of USP26 is used to investigate whether USP26 is a target of miR-203.
The effect of the increase in miR-203 using MIR203A / 5P mimic the USP26 and Snail1 in HET-1A, Kyse150 and TE-1 cell line is done by using western blot analysis of the stability of protein-based and cycloheximide. Modulating effect of miR-203 in Kyse150 and TE-1 cell lines in vitro pro-metastatic effect was analyzed by test invasion, early test wound healing, and chemosensitivity to 5-fluorouracil (5-FU). In vivo lung metastasis assay was used to study the effect of modulation of miR-203 in cells Kyse150.
Targeting deubiquitinating enzyme USP26 by microRNA-203 regulates Snail1’s pro-metastatic functions in esophageal cancer
Results: The mRNA SNAI1 and HSA / MIR203 higher and lower, respectively, in patients with EC compared with tumor-adjacent normal tissue. No changes USP26 mRNA expression was observed in this dataset. MIR / 203 expression was downregulated while the protein expression of both Snail1 and USP26 higher EC Kyse150 cell lines and TE-1 compared to normal esophageal cell lines HET-1A. USP26 is estimated as a potential target of miR-203 by TargetScan Release 2.0. Reporter tests confirmed USP26 as a target of miR-203 in the EC cell line.
Transfection of cell lines with MIR203 imitating EC decreased USP26 protein expression and protein stability Snail1 showed miR-203’s ability to regulate protein levels Snail1 through USP26. exogenous increase in miR-203 in cell lines significantly inhibited Snail EC-1 mediated in vitro function of pro-metastatic invasion, wound healing, and increased chemosensitivity to 5-FU. Finally, overexpression of miR-203 inhibited in vivo lung metastasis Kyse150 cells, which reversed following overexpression of USP26, suggesting a direct role of the regulation of miR-203-mediated metastasis USP26 in the development of EC.
Conclusion: Cumulatively, these results establish an important mechanism whereby a decrease in the expression of miR-203 potentiates the development of metastases in the EC through USP26-mediated stabilization Snail1. Therefore, miR-203 can serve as a biomarker of metastasis in EC and is a potential target for therapeutic intervention in the EC.
Dog Pancreas Specific Transcription Factor 1a ELISA kit
Description: A competitive ELISA for quantitative measurement of Canine Pancreas Specific Transcription Factor 1a in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Dog Pancreas Specific Transcription Factor 1a ELISA kit
Description: A competitive ELISA for quantitative measurement of Canine Pancreas Specific Transcription Factor 1a in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Dog Pancreas Specific Transcription Factor 1a ELISA kit
Description: A competitive ELISA for quantitative measurement of Canine Pancreas Specific Transcription Factor 1a in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: Pancreas tissue array with pancreas cancer tissue, including TNM, clinical stage and pathology grade, 80 cases/80 cores, replaced by PAN801a
Multiple tumor of pancreas and pancreas tissue array
Description: Pancreas tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.
Description: Pancreas tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.
Description: Pancreas tumor tissue lysate was prepared by homogenization in lysis buffer (10 mM HEPES pH7.9, 1.5 mM MgCl2, 10 mM KCl, 1 mM ethylenediaminetetraacetic acid, 10% glycerol, 1% NP-40, and a cocktail of protease inhibitors). Tissue and cell debris was removed by centrifugation. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.
Description: Human pancreas tissue lysate was prepared by homogenization using a proprietary technique. The tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The human pancreas tissue total protein is provided in a buffer including HEPES (pH7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, Sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the pancreas tissue pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The pancreas tissue is then Western analyzed by either GAPDH or β-actin antibody, and the expression level is consistent with each lot.
Description: Human pancreas tissue membrane protein lysate was prepared by isolating the membrane protein from whole tissue homogenates using a proprietary technique. The human pancreas tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The membrane protein is provided in a buffer including HEPES (pH 7.9), MgCl2, KCl, EDTA, Sucrose, Glycerol, sodium deoxycholate, NP-40, and a cocktail of protease inhibitors. For quality control purposes, the isolated pancreas tissue membrane protein pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The isolated pancreas tissue membrane protein is then Western analyzed by either GAPDH or β-actin antibody to confirm there is no signal or very weak signal.
Description: microRNA-21-IN-2 is a potential miR-21 inhibitor with an AC50 value of 3.29 μM. microRNA-21-IN-2 can be used for the research of cancer[1].
Description: microRNA-21-IN-3 (compound 45) can specifically bind to the precursor of oncogenic and pro-inflammatory microRNA-21 with medium nanomolar affinity, reduce cancer cell proliferation and miR-21 levels, and can be used in antitumor research[1].
Description: microRNA-21-IN-1 (compound 7A) is an efficient microRNA inhibitor. microRNA-21-IN-1 has antiproliferative activity against Hela and HCT-116 cells with IC50s of 5.5 μM and 2.8 μM respectively, as well as promotes apoptosis of Hela cells. microRNA-21-IN-1 upregulates the expression of microRNA-21 downstream functional targets (PTEN, EGR1 and SLIT2). microRNA-21-IN-1 can be used for researching anticancer[1].
Description: Human pancreas tissue cytoplasmic protein lysate was prepared by isolating the cytoplasmic protein from whole tissue homogenates using a proprietary technique. The human pancreas tissue was frozen in liquid nitrogen immediately after excision and then stored at -70°C. The cytoplasmic protein is provided in a buffer including HEPES (pH 7.9), MgCl2, KCl, EDTA, Sucrose, glycerol, and a cocktail of protease inhibitors. For quality control purposes, the isolated pancreas tissue cytoplasmic protein pattern on SDS-PAGE gel is shown to be consistent for each lot by visualization with coomassie blue staining. The isolated pancreas tissue cytoplasmic protein is then Western analyzed by GAPDH antibody, and the expression level is consistent with each lot.
Description: DNase I (EC 3.1.21.1) is an enzyme that degrade DNA, it plays a key role in the cleavage of extracellular DNA is crucial for limiting the inflammatory response and maintaining homeostasis. Exogenous deoxyribonuclease shows beneficial effects in inflammatory diseases and cancer[1].
Description: Ribonuclease A (EC 3.1.27.5) cleaves RNA 3′ to pyrimidines and actively cleaves RNA at every pyrimidine residue. Ribonuclease A catalyzes the hydrolysis of single stranded RNA in the absence of metal ions or cofactors[1][2][3].
Description: A competitive ELISA for quantitative measurement of Canine Pancreastatin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Canine Pancreastatin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Canine Pancreastatin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: Pancreas tumor tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
Description: Pancreas tumor tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
Description: Pancreas tumor tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
Description: Pancreas tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
Description: Pancreas tissue lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin. Tissue and cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The product was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol.
adverse cardiac fibrosis is involved in cardiac remodeling and heart failure, which is a major cause of heart function deteriorated. accumulative evidence has been explained that microRNAs (miRNAs) play an important role in the pathogenesis of cardiac fibrosis. However, the exact molecular mechanisms underlying the miR-144 in cardiac fibrosis is still unknown. In this study, transverse aortic constriction (TAC) and a rat model of angiotensin II (Ang II) induced cardiac fibroblasts (CF) was created to investigate the expression level of miR-144.
Douglas
